WO2006130296A2 - Systemes et procedes destines a un instrument a angle de capture eleve et a diffusion de lumiere a angles multiples (mals) - Google Patents

Systemes et procedes destines a un instrument a angle de capture eleve et a diffusion de lumiere a angles multiples (mals) Download PDF

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Publication number
WO2006130296A2
WO2006130296A2 PCT/US2006/016950 US2006016950W WO2006130296A2 WO 2006130296 A2 WO2006130296 A2 WO 2006130296A2 US 2006016950 W US2006016950 W US 2006016950W WO 2006130296 A2 WO2006130296 A2 WO 2006130296A2
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WIPO (PCT)
Prior art keywords
detector
light
target zone
particle
light beam
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Application number
PCT/US2006/016950
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English (en)
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WO2006130296A3 (fr
Inventor
Scott H. Bloom
Victor J. Chan
Steven A. Cashion
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Jmar Research, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US11/231,350 external-priority patent/US7532327B2/en
Application filed by Jmar Research, Inc. filed Critical Jmar Research, Inc.
Priority to EP06784381A priority Critical patent/EP1907820A4/fr
Publication of WO2006130296A2 publication Critical patent/WO2006130296A2/fr
Publication of WO2006130296A3 publication Critical patent/WO2006130296A3/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1468—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle
    • G01N15/147—Optical investigation techniques, e.g. flow cytometry with spatial resolution of the texture or inner structure of the particle the analysis being performed on a sample stream
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1429—Signal processing
    • G01N15/1433—Signal processing using image recognition
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
    • G01N2015/019—Biological contaminants; Fouling

Definitions

  • Embodiments of the invention relate to identifying particles, and in particular to identifying particles in a liquid using illumination incident at an angle.
  • Rapid tests include immunoassays, ATP luminescence, and fluorescent antibody fixation. Rapid tests are also by grab sample and require manipulation of the sample to 'tag' the microbes with an identifiable marker or concentrate the microbe's genetic material (DNA) for subsequent identification. " Results are normally available in 1-3 hours. These types of tests include Polymerase Chain Reaction (PCR) and Flow Cytometry.
  • PCR Polymerase Chain Reaction
  • Real time contamination warning systems are continuous warning devices that detect contaminants and provide an 'event' warning within minutes to prompt further investigation or action. CWS include laser based multi-angle light scattering (MALS) and multi-parameter chemical & particle instruments that detect water quality changes inferring potential biological contamination. Continuous, real time detection of pathogens in water surveillance was first discovered in the late 1960's and has progressed through a series of development steps until the first public field demonstration in 2002.
  • MALS laser based multi-angle light scattering
  • CWS include laser based multi-parameter chemical & particle instruments that detect water quality changes inferring
  • MALS is an acronym for "multi-angle light scattering" and is based on laser technology, photo-detection, and computer signal processing.
  • a characteristic scattering pattern is emitted.
  • the scattering pattern encompasses many features of the particle including the size, shape, internal structures (morphology), particle surface, and material composition (organic or inorganic).
  • Each type of microorganism will scatter light giving off a unique pattern called a 'bio-optical signature'.
  • Photo- detectors collect the scattered light and capture the patterns which are then sent to an on-board computer. A microorganism's bio-optical signature is then compared against known pattern classifications in the detection library for results.
  • a particle detection system uses a reflective optic comprising a curved surface to detect high angle scattered light generated by a particle in a liquid medium, when a laser beam is incident on the particle.
  • a reflective optic comprising a curved surface to detect high angle scattered light generated by a particle in a liquid medium, when a laser beam is incident on the particle.
  • the particles transit the laser beam, light is scattered in all directions and is described by MIE scattering theory for particles about the size of the wavelength of light and larger or Rayleigh Scattering when the particles are smaller than the wavelength of light.
  • the reflective optic the scattered light can be detected over angles that are greater than normally obtainable. For example, the scattered light can be measured through an angle 90°.
  • Figure 1 is a diagram illustrating an example embodiment of a particle detection system
  • Figure 2 is a diagram illustrating another example embodiment of a particle detection system
  • Figure 3 A is a picture of B. suptilis spores
  • Figure 3B and 3C are pictures illustrating example optical signatures that can be generated by the systems of figures 1 and 2 for the B. suptilis spores of figure 3 A;
  • Figure 4A is a picture of a ball of plastic spheres
  • Figure 4B and 4C are pictures illustrating example optical signatures that can be generated by the systems of figures 1 and 2 for the ball of plastic spheres of figure 4A;
  • Figures 5-7 are diagrams illustrating a technique for using illumination incident at an angle in a light scattering detection system, such as the systems of figures 1 and 2;
  • Figure 8 is a diagram illustrating an example particle detection system that implements the technique of figures 5-7 in accordance with one embodiment
  • Figure 9 is a diagram illustrating an example particle detection system that implements the technique of figures 5-7 in accordance with another embodiment
  • Figure 10 is a diagram illustrating a spectrometer ray trace for light scattered by a particle suspended in a liquid medium and reflected by a curved mirror;
  • Figure 11 is a diagram illustrating the scattered light pattern produced by the particle of figure 10;
  • Figure 12 is a graph illustrating the relative intensity of the scattered light versus the scattering angle.
  • k is 1%, 2%, 3%, 4%, 5%, ..., 50%, 51%, 52%,..., 95%, 96%, 97%, 98%, 99%, or 100%.
  • any numerical range defined by two numbers, R, as defined in the above is also specifically disclosed. It is also emphasized that in accordance with standard practice, various features may not be drawn to scale. In fact, the dimensions of the various features may be arbitrarily increased or reduced for clarity of discussion.
  • Embodiments of the present invention provide a method for realtime particle detection that uses advancements in computing power, special optics, photonics engineering, advanced signal processing, and complex algorithms, in order to provide a MALS detection system that provides simplicity, cost effectiveness, speed, and reliability.
  • the systems described in the embodiments below are analytical system using MALS where a side stream from a water source flows through a transparent flow cell. A laser directs a beam of light into the flow cell and through the water stream.
  • the water is first characterized for background interferences to distinguish foreign particles from the pathogens' signatures resulting in a custom detection library in each particular installation.
  • particles pass through the beam, the scattered light is emitted and captured by the detectors, converted to a digital signal, and finally sent to the computer's microbial library for analysis.
  • the organisms are classified within minutes.
  • the data can be transmitted to a user screen and remote communications equipment.
  • an 'alert' upon reaching a pre-set threshold level, an 'alert' can be generated and an instantaneous sample can be automatically extracted for further identification and confirmation.
  • Water, or other liquids for that matter can be monitored continuously as it passes through the flow cell at a defined rate. This provides a much higher probability of detecting and classifying microorganisms compared to intermittent grab samples. The speed and performance can be further enhanced when the 1) microbial concentration level is high, 2) the water, or liquid, is of high 'clarity' or purity, 3) microorganisms match defined bio-optical signatures in the library versus an 'unknown', and 4) the particles are of larger size, e.g., >1 micron, giving distinct scattering patterns.
  • the system can categorize it as an 'unknown' and still provide an 'alert' if a certain threshold level is reached.
  • FIG. 1 is a diagram illustrating an example particle detection system configured in accordance with one embodiment of the systems and methods described herein.
  • System 100 comprises a light source 102 configured to provide illumination 104 to a target area 108.
  • target area 108 is within a fluid cell 106.
  • Water intended to be interrogated for various particles, or microorganisms can flow through flow cell 106, e.g., in a downward direction as indicated.
  • Illumination 104 will encounter particles in target zone 108, which will cause the illumination to scatter in a manner different than the illumination transmitted through the surrounding fluid medium.
  • System 100 can also comprise an optical system 124.
  • Optical system 124 can comprise several elements.
  • optical system 124 can comprise a lens, or lens system 112 as well as an optical element 114.
  • the system 100 can also comprise a detector, detector system, or detector array 116, which can be interfaced with a processing system 118.
  • Light source 102 can be configured to deliver a structured light pattern, or illumination.
  • light source 102 can be, e.g., a coherent light source, such as a laser.
  • light source 102 can comprise a single light source, such as a single laser, or a plurality of light sources, such as a plurality of lasers.
  • the wavelength of the light source can be at a fixed wavelength. Alternatively, when multiple light sources are used, the light sources can have several discrete wavelengths.
  • light source 102 can be a laser configured to produce a laser beam 104.
  • laser beam 104 strikes a particle within target area 108, the particle will cause the beam to scatter in a pattern that is different than the pattern produced due to beam 104 traveling through the water flowing in flow cell 106.
  • Optical system 124 can be configured to then pick up the scattered light and direct it onto detector 116.
  • Detector 116 can actually be a plurality of detectors, such as a plurality of detectors arrayed in different positions around target area 108.
  • detector 116 can comprise an array of photo detectors.
  • detector 116 can actually comprise a linear array of photo detectors configured to detect the scattered light and generate an electrical signal having an amplitude corresponding to the amplitude of the detected light.
  • a Charge Coupled Device CCD
  • CCDs are readily available with thousands of pixels, wherein each pixel can form an individual photo detector.
  • a 2 dimensional array of photodiodes or avalanche photodiodes of 64, 128, 256, or 512 total pixels can be used to increase the total dynamic range of the detector as compared to a CCD.
  • Detector 116 can be configured to generate an electrical signal, or signals, reflective of the light pattern incident on detector 116. The signals can then be provided to processing system 118 for further analysis. As described above, processing system 118 can convert the signals into a pattern using various algorithms 122. Processing system 118 can also comprise the memory configured to store a plurality of optical signatures, or patterns 120 that are associated with various particles, or microorganisms of interest.
  • processing system can compare the pattern generated using algorithms 122 to one of the stored patterns 120 in order to identify particles within target zone 108.
  • algorithms 122 and patterns 120 can be used to determine many features of particles being identified within target zone 108, e.g., including the size, shape, internal structures or morphology, particle surface, and material composition, i.e., organic or inorganic.
  • certain embodiments can use Multiple Analysis Of Variance (MANOVA) algorithms, neuro networks, simulated and annealing, algorithm independent machine learning, physiologic, grammatical methods, and other algorithmic techniques for pattern generation and recognition.
  • MANOVA Multiple Analysis Of Variance
  • a spherical lens (not shown) completely suiTounding the flow cell, except for the flow cell inlet and outlet, can be placed at the interface of flow cell 106 in order to allow light scattered at any angle to the lens to pass through the lens to optical system 124.
  • a spherical lens increases the complexity and cost of system 100.
  • Light passing through target zone 108 along the optical axis of beam 104 will generally be of a much greater intensity than that of the scattered light beams. The intensity of the beam along the optical axis can be so great that it can essentially prevent, or degrade detection of the scattered light beams.
  • a beam stop 110 can be included in order to deflect beam 104 and prevent it from entering optical system 124 and being detected by detector 116.
  • the light scattered by a particle within target zone 108 can enter optical system 124, which can comprise an optical element 114.
  • Optical element 114 can be configured to direct the scattered light onto detector 116.
  • optical element 114 can be configured in such a way that it can direct light traveling along a given path to an appropriate position on detector 116 or to an appropriate detector within an array of detectors comprising detector 116.
  • optical element 114 can be a holographic optical element constructed so that each refracting section refracts, or redirects light from one of the scattered paths so that it falls on the correct location of detector 116.
  • optical element 114 can comprise a zone plate lens that can be configured to map the distance from the central optical access to a unique mapping that is useful for high speed scanning.
  • the scattered light may need to be collimated after it passes through target zone 108.
  • a converging lens 112 can be included in optical system 124.
  • a converging lens can be configured to reduce the angle spread for the various scattered light rays.
  • a converging lens can be configured to collimate or converge the spread light rays.
  • some other optical device can be used to collimate the scattered light rays. It will also be apparent, that certain embodiments may not need an optical lens 112, i.e., collimation may not be necessary depending on the embodiment.
  • optical system 124 may or may not contain an optical lens 112, or a collimator, as required by the specific implementation.
  • detector 116 can actually comprise a plurality of detectors such as a linear detector array or 2 dimensional array such as a Charge Coupled Device (CCD) or for better dynamic range, a 2 dimensional array of photodiodes or avalanche photodiodes.
  • detector 116 can actually comprise a linear photo diode camera, e.g., a 128-pixel linear photo diode camera.
  • an square array of photodiodes may be used for detector 116.
  • an array of photodiodes arranged in segmented concentric circles may be employed for detector 116.
  • optical element 114 will be selected so as to complement detector 116 by directing the scattered light rays onto the appropriate pixel, or a section of detector 116; however, in certain embodiments, optical element 114 may not be needed. For example, in certain embodiments, the scattered light rays are incident directly onto detector 116.
  • FIG. 2 is a diagram of a particle detection system 200 that does not include an optical element.
  • system 200 comprises a light source 202, such as a laser, that produces a beam 204 that is incident on particles in target zone 208 within a fluid flowing in flow cell 206.
  • the particles scatter beam 204 and the scattered beams are then incident directly on a detector 212.
  • Detector 212 then produces electrical signals based on the incident scattered light rays and provides the electrical signals to processing system 214.
  • Processor system 214 can, like processing system 118, be configured to generate a pattern from the electrical signals using algorithms 218 and compare them against stored patterns 216 in order to identify particles within target zone 208.
  • a beam stop 210 is still required to reflect the light ray traveling along the optical axis.
  • detector 212 can comprise a 64- pixel detector array, while in other embodiments, detector 212 can comprise a 128-pixel detector array. In certain embodiments, it can be preferred that detector 212 comprise a 256-pixel detector. Arrays larger than 256-pixels can be utilized in the present invention at a penalty of increasing cost and complexity. It should also be noted, that detector 212 can comprise conditioning amplifiers, multiplex switches, an Analog-to-Digital Converter (ADC) configured to convert analog signals produced by the detector pixel elements into digital signals that can be passed to processing system 214. An example embodiment of a detector is described in more detail below with respect figure 14.
  • ADC Analog-to-Digital Converter
  • system 200 can include telescoping optics (not shown) in order to collimate the scattered light rays if necessary.
  • each type of particle, or microorganism will scatter light giving off a unique pattern called an optical signature, or bio-optical signature.
  • a detector such as detector 212, can collect the scattered light and capture the patterns. Electrical signals representative of the pattern can then be provided to a processing system such as processing system 214.
  • Figures 3 and 4 illustrate example optical signatures for two different types of particles.
  • Figure 3A is a picture illustrating subtilis spores, a microorganism.
  • Figures 3B and 3 C are pictures illustrating the optical signature associated with the subtilis spores of figure 3 A.
  • Figure 4A is a picture illustrating a ball of plastic spheres.
  • Figures 4B and 4C are diagrams illustrating the optical signature for the ball of plastic spheres in figure 4A.
  • the optical signatures, or patterns, of figures 3A-3B and 4A -4B which can be produced using, e.g., algorithms 218, can be compared to patterns stored within the processing system.
  • vector kj can be used to represent the illumination.
  • illumination incident along vector k; encounters particle 502 it will be scattered through a sphere of 360 degrees but only detected through a range of angles up to ⁇ .
  • a scattered light ray at the outer edge of the detector range can be represented by vector k s .
  • the detector will be able to see light scattered through a greater range of angles.
  • the scattered light rays will be measured through an angle of 2 ⁇ .
  • objective 500 can collect scattered light rays scattered through twice the angle as compared to the system in figure 5.
  • the resolution of the system illustrated in figure 6 would be twice that of the system illustrated in figure 5.
  • Figure 7 is a diagram illustrating that the same effect can be achieved using a plurality of incident beams 508 that include beams incident at an angle from above and below the optical axis 504. Switching on or off the individual laser beams can provide additional multiple angles without having to provide additional detectors. If the switching is fast enough compared to the transit of the particle through the beam, then the additional angles can be obtained for the same particle.
  • objective 500 in figures 5-7 can be a zone plate as well as another conventional optical element, including a holographic optical element.
  • FIGs 8 and 9 illustrate that the technique depicted in figures 6 and 7 could be achieved by altering the position of the optical detector or by configuring the light source so that the illumination is incident at an angle upon the target zone.
  • figure 8 is a diagram illustrating an example particle detection system 800 in which an optical detector 812 has been repositioned so as to capture scattered light rays scattered to an angle 2 ⁇ .
  • a light source 802 such as a laser, produces a beam 804 that is incident on particles within target zone 808.
  • a beam stop 810 can still be required within system 800 to deflect the beam traveling along the optical axis.
  • system 800 can comprise a processing system, but that such system is not illustrated for simplicity.
  • Figure 9 is a diagram illustrating an example particle detection system 900 in which optical source 902 is configured such that beam 904 is incident upon target zone 908 at an angle equal to or greater than the critical angle defined by the phenomenon or total internal reflection.
  • optical source 902 is configured such that beam 904 is incident upon target zone 908 at an angle equal to or greater than the critical angle defined by the phenomenon or total internal reflection.
  • the beam 904 is internally reflected within flow cell 906, and thus a beam stop is not required. This can lower the cost and complexity of system 900 and can, therefore, be preferable.
  • system 900 can comprise a processing system, but that such system is not illustrated for simplicity.
  • TIR Total Internal Reflection
  • TIR can occur at high to low indexes of refraction interfaces within the optics of the instrument, or system being used to observe or collect the scattered light, e.g., the interface between flow cell 106 and the external atmosphere.
  • a second surface curved mirror reflecting optic can be used to collect and reflect the light.
  • Such an optic can allow easy capture of light angles up to 90° for all azimuthal angles, when the sample is index coupled with the non-reflecting surface of the collection optic.
  • Such an optic can prevent TIR issues at angles greater than approximately 40°.
  • Figure 10 is a diagram illustrating a scatterometer ray trace for light scattered by a particle 1002 and collected using a second surface curved mirror 1004.
  • light reflected through an angle of 60° by the reflective surface of mirror 1004 corresponds to light scattered through an angle of 90° by object 1002.
  • the scattered light 1008 passes by beam stop 1006, which is configured to reflect the high intensity light traveling along the beam axis. Scattered light can then be incident on a detector surface 1010, such as a CCD.
  • Figure 11 is a diagram illustrating a pattern produced by scattered light 1008 incident on detector 1010.
  • the pattern depicted in figure 11 corresponds to the diffraction pattern generated by a sphere comprising a diameter of approximately 8 microns.
  • Line 1102 is drawn along the laser polarization axis.
  • Beam stop 1006 reflects light along the beam axis.
  • Figure 12 is a graph illustrating the relative intensity of scattered light versus the scatter angle for the pattern of figure 11. As can be seen, light scattered through an angle of 90° can be detected using optic 1004.
  • a reflective optic such as optic 1004 can be included in systems such as systems 100 and 200.
  • An optic such as optic 1004 can be included in place of, or in addition to other optics with in the system. This can increase the angle ⁇ through which scattered light can be collected and detected.
  • systems 100 and 200 are just examples of the types of systems that can make use of a second surface curved mirror for collecting and detecting high angel scattered light as describe above. Accordingly the embodiments described with respect to figures 10-12 should not be seen as limited to implementation in systems such as systems 100 and 200.
  • figure 13 is a diagram illustrating a system 1300 configured to collect light scattered by a particle and reflected by a curved reflective optic as described above.
  • System 1330 comprises a laser 1302 configured to generate a laser beam 1304.
  • Beam 1304 can be directed at a 45 degree reflective silver prism 1306, which can cause beam 1304 to go through interface optic 1308, flow cell 1310, and reflective optic 1312 through unsilvered area 1314 on reflective optic 1312.
  • silver prism 1306 and unsilvered area 1314 on reflective optic 1312 allow beam 1304 to be removed from the desired signal, much as beamstop 1006 does in alternative embodiments.
  • Interface optical element 1308 can be a separate element optically coupled to flowcell 1310 with a coupling medium, or integral to the design of the flow cell 1310.
  • Reflective optical element 1312 can also be a separate element optically coupled to flowcell 1310 with a coupling medium or integral to flowcell 1310. The scattered radiation pattern produced by an object in flowcell 1310 is reflected by reflective optical element 1312. The reflected light then falls on the 2- dimensional photo detector array 1316.
  • Figure 14 is a diagram illustrating an example detector system
  • system 1400 such as detector 212 or a system including array 1316.
  • system 1400 comprises a 256 -pixel detector packaged array 1402 removably attached to a signal conditioning and digitizing board 1430.
  • Board 1430 can comprise signal conditioning amplifiers 1406 and 1408, multiplex analog switches 1410, a 14-bit Analog to Digital Converter (ADC) 1416, a microcontroller 1418, and a USB2.0 communications chip 1420.
  • ADC Analog to Digital Converter
  • microcontroller 1418 a microcontroller 1418
  • USB2.0 communications chip 1420 USB2.0 communications chip

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
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Abstract

Un système de détection de particules utilise une optique réfléchissante comprenant une surface incurvée pour détecter une lumière diffusée à angle élevé, générée par une particule dans un milieu liquide, le faisceau laser étant incident à la particule. Lorsque les particules transitent par le faisceau laser, la lumière est diffusée dans toutes les directions et est décrite par une théorie de diffusion MIE pour les particules en ce qui concerne la taille de la longueur d'onde de la lumière (ou plus large) ou par la diffusion de Rayleigh lorsque les particules sont plus petites que la longueur d'onde de la lumière. L'utilisation de l'optique réfléchissante permet de détecter la lumière diffusée à des angles plus grands que les angles obtenus normalement.
PCT/US2006/016950 2005-05-02 2006-05-02 Systemes et procedes destines a un instrument a angle de capture eleve et a diffusion de lumiere a angles multiples (mals) WO2006130296A2 (fr)

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EP06784381A EP1907820A4 (fr) 2005-05-02 2006-05-02 Systemes et procedes destines a un instrument a angle de capture eleve et a diffusion de lumiere a angles multiples (mals)

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US67673005P 2005-05-02 2005-05-02
US60/676,730 2005-05-02
US11/231,350 2005-09-19
US11/231,350 US7532327B2 (en) 2004-09-17 2005-09-19 Systems and methods for detecting scattered light from a particle using illumination incident at an angle

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US7564551B2 (en) 2009-07-21
EP1907820A4 (fr) 2011-07-06
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US20060256333A1 (en) 2006-11-16
EP1907820A2 (fr) 2008-04-09

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